RESUMO
The study of venom components is an important step toward understanding the mechanism of action of such venoms and is indispensable for the development of new therapies. This work aimed to investigate the venom of Lachesis muta rhombeata and evaluate enzymes related to its toxicity. Phospholipase A2 (PLA(2)), L-amino acid oxidase (LAAO), and proteinase activities were measured, and the molecular weights were estimated. We found the venom to contain one PLA(2) (17 kDa), one LAAO (132 kDa), and three serine proteinases (40, 31, and 20 kDa). Although only serine proteinases were observed in the zymogram, metalloproteinases were found to contribute more to the total proteolytic activity than did serine proteinases. The work confirmed the presence of highly active enzymes; and, moreover, we proposed a novel method for confirming the presence of LAAOs by zymography. We also suggested a simple step to increase the sensitivity of proteinase assays.
Assuntos
L-Aminoácido Oxidase/química , Peptídeo Hidrolases/química , Fosfolipases A2/química , Venenos de Víboras/enzimologia , Viperidae , Animais , Caseínas/química , Embrião de Galinha , Eritrócitos/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Leucina/química , Camundongos , Peso Molecular , Oxirredução , Fosfolipases A2/farmacologia , Inibidores de Proteases/química , Proteólise , Soroalbumina Bovina/química , Venenos de Víboras/química , Venenos de Víboras/farmacologiaRESUMO
BACKGROUND: The aim of this work was to study the new bone tissue formation after bone morphogenetic protein type 2 (rhBMP-2) and P-1 application, using 5 and 10 µg of each, combined to a material carrier, in critical bone defects. METHODS: It was used 70 Wistar rats (male, â¼250 g) that were divided in 10 groups with seven animals on each. Groups are the following: critical bone defect only, pure monoolein gel, 5 µg of pure P-1, 5 µg of pure rhBMP-2, 5 µg of P-1/monoolein gel, 5 µg of rhBMP-2/monoolein gel, 10 µg of pure P-1, 10 µg of pure rhBMP-2, 10 µg of P-1/monoolein gel, 10 µg of rhBMP-2/monoolein gel. Animals were sacrificed after 4 weeks of the surgical procedure and the bone samples were submitted to histological, histomorphometrical, and immunohistochemical evaluations. RESULTS: Animals treated with pure P-1 protein, in both situations with 5 µg and 10 µg, had no significant difference (P > 0.05) for new bone formation; other groups treated with 10 µg were statistically significant (P < 0.05) among themselves and when compared with groups in which it was inserted the monoolein gel or critical bone defect only (P < 0.05). In the group involving the 10 µg rhBMP-2/monoolein gel association, it was observed an extensive bone formation, even when compared with the same treatment without the gel carrier. CONCLUSION: Using this experimental animal model, more new bone tissue was found when it was inserted the rhBMP-2, especially when this protein was combined to the vehicle, and this process seems to be dose dependent.
Assuntos
Proteína Morfogenética Óssea 2/farmacologia , Regeneração Óssea/efeitos dos fármacos , Látex/química , Proteínas de Plantas/farmacologia , Fator de Crescimento Transformador beta/farmacologia , Animais , Proteína Morfogenética Óssea 2/administração & dosagem , Relação Dose-Resposta a Droga , Portadores de Fármacos/administração & dosagem , Glicerídeos/administração & dosagem , Hevea/química , Humanos , Imuno-Histoquímica , Masculino , Modelos Animais , Proteínas de Plantas/administração & dosagem , Ratos , Ratos Wistar , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Crânio/efeitos dos fármacos , Crânio/lesões , Fator de Crescimento Transformador beta/administração & dosagemRESUMO
This in vivo study evaluated the osteogenic potential of two proteins, recombinant human bone morphogenetic protein-2 (rhBMP-2) and a protein extracted from natural latex (Hevea brasiliensis, P-1), and compared their effects on bone defects when combined with a carrier or a collagen gelatin. Eighty-four (84) Wistar rats were divided into two groups, with and without the use of collagen gelatin, and each of these were divided into six treatment groups of seven animals each. The treatment groups were: (1) 5 microg of pure rhBMP-2; (2) 5 microg of rhBMP-2/monoolein gel; (3) pure monoolein gel; (4) 5 microg of pure P-1; (5) 5 microg of P-1/monoolein gel; (6) critical bone defect control. The animals were anesthetized and a 6 mm diameter critical bone defect was made in the left posterior region of the parietal bone. Animals were submitted to intracardiac perfusion after 4 weeks and the calvaria tissue was removed for histomorphometric analysis. In this experimental study, it was concluded that rhBMP-2 allowed greater new bone formation than P-1 protein and this process was more effective when the bone defect was covered with collagen gelatin (P < 0.05).
Assuntos
Doenças Ósseas/tratamento farmacológico , Proteína Morfogenética Óssea 2/farmacologia , Regeneração Óssea/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteínas de Plantas/farmacologia , Proteínas Recombinantes/farmacologia , Animais , Proteína Morfogenética Óssea 2/uso terapêutico , Regeneração Óssea/fisiologia , Osso e Ossos/citologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Fraturas Ósseas/tratamento farmacológico , Fraturas Ósseas/patologia , Fraturas Ósseas/fisiopatologia , Hevea/química , Humanos , Látex/química , Masculino , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Extratos Vegetais/uso terapêutico , Proteínas de Plantas/uso terapêutico , Ratos , Ratos Wistar , Proteínas Recombinantes/uso terapêutico , Resultado do TratamentoRESUMO
Objetivos: Avaliar clinicamente a eficácia do tratamento dos diferentes tipos de úlceras de perna com a biomembrana de látex natural e verificar os efeitos adversos mais comuns do tratamento. Pacientes e métodos: foram avaliados 21 pacentes portadores de úlceras de perna do ambulatório de dermatologia HU-UFJF, selecionados aleatoriamente e submetidos ao protocolo. Os curativos constituiam-se de: lavagem das feridas com soro fisiológico 0,9%, aplicação da biomembrana sobre o fundo da úlcera e oclusão realizados no domicilio, pelo pacienteem dias alternados, sendo acompanhado semanalmente no ambulatório. Resultados: a média de idade dos pacientes foi de 64.5 anos, sendo 29% do sexo masculino e 71% feminino. A insuficiência venosa isoladamente foi encontrada em 33,3% dos pacientes e associada à HAS em 52.4%. As úlceras foram classificadas como venosas em 57.6%, hipertensivas em 33.6% e outras em 9.6% com o tempo médio de duração de 6,15 anos. Sinais clínicos de granulação foram evidenciados já na primeira semana em 100% dos casos. Sinais de reepitelização foram observados em 52,4% dos pacientes, num tempo médio de tratamento de 41,4 dias. O surgimento ou aumento da dor foi o efeito adverso mais relatado em 57% das úlceras hipertensivas e em 25% das venosas. O aumento das úlceras foi causa de suspensão da biomembrana em 57% das hipertensivas e em 8% das venosas, sendo que esta apresentou complicação por erisipéla.
Assuntos
Humanos , Masculino , Feminino , Tecido de Granulação , Úlcera da Perna , Látex/uso terapêutico , Polilisina , Úlcera da Perna/prevenção & controle , CicatrizaçãoRESUMO
BACKGROUND: Clinical symptoms and outcomes of uraemic patients ingesting star fruit are quite variable and may progress to death. The purpose of the present report was to discuss the neurotoxic effects of star fruit intoxication in uraemic patients and to present the efficacy of different therapeutic approaches. METHODS: We studied a total of 32 uraemic patients who had ingested star fruit. Before the intoxication episodes, 20 patients were on regular haemodialysis, eight were on peritoneal dialysis and four were not yet undergoing dialysis. Two patients were analysed retrospectively from their charts, 17 were directly monitored by our clinic and 13 were referred by physicians from many areas throughout the country, allowing us to follow their outcome from a distance. Intoxicated patients were given different therapeutic approaches (haemodialysis, peritoneal dialysis and supportive treatment), and their outcomes were analysed. RESULTS: The most common symptoms were persistent and intractable hiccups in 30 patients (93.75%), vomiting in 22 (68.7%), variable degrees of disturbed consciousness (mental confusion, psychomotor agitation) in 21 (65.6%), decreased muscle power, limb numbness, paresis, insomnia and paresthesias in 13 (40.6%) and seizures in seven (21.8%). Patients who were promptly treated with haemodialysis, including those with severe intoxication, recovered without sequelae. Patients with severe intoxication who were not treated or treated with peritoneal dialysis did not survive. CONCLUSIONS: Haemodialysis, especially on a daily basis, is the ideal treatment for star fruit intoxication. In severe cases, continuous methods of replacement therapy may provide a superior initial procedure, since rebound effects are a common event. Peritoneal dialysis is of no use as a treatment, especially when consciousness disorders ensue.